Evidence map›Paper›PMID 41339334›Full record

ArticleNature communications2025

A BRET biosensor for measuring uncompetitive engagement of PRMT5 complexes in cells.

Elisabeth M Rothweiler, Ani Michaud, Jakub Stefaniak, Usha Singh, Brynwood B Mikulsky, James D Vasta, Michael T Beck, Jennifer Wilkinson, Jennifer A Ward, Catherine M Rogers and 9 more

Abstract read
PubMed Full text at PMC Publisher
In one paragraph

Article in Nature communications, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 2 papers.

0numbers the graph read from it
0cells of the map it votes in
2citing papers in PubMed
–field-weighted citation impact
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

2 citing papers in PubMed.

  1. Review
  2. Review
4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

19 authors.

Elisabeth M Rothweiler *Centre for Medicines Discovery, Nuffield Department of Medicine, University of Oxford, Old Road Campus, Oxford, UK.
Ani Michaud *Promega Corporation, Madison, WI, USA.
Jakub StefaniakCentre for Medicines Discovery, Nuffield Department of Medicine, University of Oxford, Old Road Campus, Oxford, UK.
Usha SinghCentre for Medicines Discovery, Nuffield Department of Medicine, University of Oxford, Old Road Campus, Oxford, UK.
Brynwood B MikulskyPromega Corporation, Madison, WI, USA.
James D VastaPromega Corporation, Madison, WI, USA.
Michael T BeckPromega Corporation, Madison, WI, USA.
Jennifer WilkinsonPromega Corporation, Madison, WI, USA.ORCID http://orcid.org/0009-0002-1994-715X
Jennifer A WardCentre for Medicines Discovery, Nuffield Department of Medicine, University of Oxford, Old Road Campus, Oxford, UK.
Catherine M RogersCentre for Medicines Discovery, Nuffield Department of Medicine, University of Oxford, Old Road Campus, Oxford, UK.
Esra BalıkçıCentre for Medicines Discovery, Nuffield Department of Medicine, University of Oxford, Old Road Campus, Oxford, UK.ORCID http://orcid.org/0009-0000-9160-5790
Jeppe Tranberg-JensenCentre for Medicines Discovery, Nuffield Department of Medicine, University of Oxford, Old Road Campus, Oxford, UK.
Jesper S HansenCentre for Medicines Discovery, Nuffield Department of Medicine, University of Oxford, Old Road Campus, Oxford, UK.
Peter LoppnauStructural Genomics Consortium, University of Toronto, Toronto, ON, Canada.
Adrian WhittyDepartment of Chemistry, Boston University, Boston, MA, USA.ORCID http://orcid.org/0000-0002-0331-4369
Paul E BrennanCentre for Medicines Discovery, Nuffield Department of Medicine, University of Oxford, Old Road Campus, Oxford, UK.
Peter J TongeCenter for Advanced Study of Drug Action, Department of Chemistry, Stony Brook University, Stony Brook, NY, USA. peter.tonge@stonybrook.edu.ORCID http://orcid.org/0000-0003-1606-3471
Matthew B RobersPromega Corporation, Madison, WI, USA. matt.robers@promega.com.ORCID http://orcid.org/0000-0001-8505-5227
Kilian V M HuberCentre for Medicines Discovery, Nuffield Department of Medicine, University of Oxford, Old Road Campus, Oxford, UK. kilian.huber@cmd.ox.ac.uk.ORCID http://orcid.org/0000-0002-1103-5300

Funding

Mechanism of Slow Onset Enzyme Inhibition and Translation to Time-Dependent Drug ActivityR35GM149297 · NIGMS · STATE UNIVERSITY NEW YORK STONY BROOK · PI PETER J TONGE · 2023 to 2026
$1.6M
Innovative Medicines Initiative (IMI) 875510NIGMS NIH HHS R35 GM149297
6 · The paper itself

Abstract

Protein arginine methyl transferase 5 (PRMT5) plays a global role in cell physiology and is an established therapeutic target in cancer. In approximately 10-15% of human cancers, deletion of the methylthioadenosine phosphorylase (MTAP) gene results in accumulation of methylthioadenosine (MTA), exposing a synthetic lethality and opportunity for precision medicine by selective targeting of PRMT5 in this context. Reported small molecule PRMT5 inhibitors engage either cosubstrate S-adenosyl methionine (SAM) or peptide-substrate pockets through diverse mechanisms. A subset of chemotypes demonstrate uncompetitive engagement with SAM or its inhibitory metabolic precursor, MTA. Although uncompetitive engagement can be evaluated in cell-free systems, no methods exist to directly assess this in cells. Here, we describe the development of a fluorescent probe that acts as a dynamic BRET biosensor of the intracellular SAM/MTA pool that overcomes the current limitations of competitive binding analyses. Using this biosensor, we evaluate a range of diverse PRMT5 inhibitors to mechanistically characterize and quantify uncompetitive target engagement as well as ternary complex formation at PRMT5-SAM and PRMT5-MTA complexes in live cells, enabling direct insights into drug mechanism-of-action and metabolite-dependent responses of inhibitors.

Indexed as

Biosensing TechniquesProtein-Arginine N-MethyltransferasesDeoxyadenosinesFluorescent DyesHEK293 CellsHumansPurine-Nucleoside PhosphorylaseS-AdenosylmethionineThionucleosides5'-methylthioadenosine5'-methylthioadenosine phosphorylaseDeoxyadenosinesFluorescent DyesPRMT5 protein, humanProtein-Arginine N-MethyltransferasesPurine-Nucleoside PhosphorylaseS-AdenosylmethionineThionucleosides

Identifiers

PMID41339334
PMCPMC12675575

What OpenQuestion holds

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Registered trials

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.